Poly (ADP-Ribose) Polymerase I (PARP-1) is a first responder to DNA damage and participates in the regulation of gene expression. The interaction of PARP-1 with chromatin and DNA is complex and involves at least two different modes of interaction. In its enzymatically inactive state, PARP-1 binds native chromatin with similar affinity as it binds free DNA ends. Automodification of PARP-1 affects interaction with chromatin and DNA to different extents. Here we describe a series of biochemical and biophysical techniques to quantify and dissect the different binding modes of PARP-1 with its various substrates. The techniques listed here allow for high throughput and quantitative measurements of the interaction of different PARP-1 constructs (inactive and automodified) with chromatin and DNA damage models.
CITATION STYLE
Chassé, M. H., Muthurajan, U. M., Clark, N. J., Kramer, M. A., Chakravarthy, S., Irving, T., & Luger, K. (2017). Biochemical and biophysical methods for analysis of poly(ADP-ribose) polymerase 1 and its interactions with chromatin. In Methods in Molecular Biology (Vol. 1608, pp. 231–253). Humana Press Inc. https://doi.org/10.1007/978-1-4939-6993-7_16
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